JP4468617B2 - Method for concomitant administration of anticancer agents and anticancer agents that can be used in combination - Google Patents

Description

【００１２】
本発明者らは、シスプラチン、カルボプラチン、５−ＦＵ、テガフール、カルモフール、ドキシフルリジン、ウラシル、イリノテカン、アドリアマイシン、エトポシド、マイトマイシン、ミトキサントロン、ブレオマイシン、ビンクリスチン及びビンデシンの計１５種類の既知の抗癌剤（併用剤）と１−ＯＨＰ（標的剤）との併用投与による腫瘍細胞の死滅効果の測定を試みる。該併用に際しては単に抗癌剤の併用だけでなく、投与方法（同時投与、追加投与）及び投与量（多量投与、少量投与）についても検討する。
【００１３】
各組合せにおける腫瘍細胞の死滅効果の評価に際しては、対照（抗癌剤を使用しない場合）の死滅効果を差引き、純粋な併用剤と１−ＯＨＰの併用による死滅効果を算出する。
併用効果は、後述の式（１）を使用して算出し、この式により算出される値（％）が、８５〜１１５％がある場合には相加効果があり、８５％未満の場合には相加効果以下、１１５％を越える場合には相乗効果を有するものと、評価する。±１５％は信頼限界つまり誤差である。
【００１４】
その結果、下記に詳述するように、シスプラチン、５−ＦＵ、イリノテカン、アドリアマイシン、エトポシド、マイトマイシン、ミトキサントロン及びブレオマイシンの計８種類についてはその殆どについて投与方法及び投与量にかかわらず、相加効果あるいは相乗効果が得られ、特にシスプラチンと５−ＦＵの併用における効果は著しく、５−ＦＵと１−ＯＨＰの組合せが最高である。一方ビンクリスチン及びビンデシンの計２種類については良くても相加効果が現れるのみの組合せが多い。
【００１５】
つまり、シスプラチンと１−ＯＨＰの併用投与では、シスプラチンを同時投与した場合、及び１−ＯＨＰをシスプラチンより先行投与した全ての組合せで相乗効果が得られ、条件によっては期待値（相加効果）の２倍の腫瘍細胞の死滅効果が現れる。
５−ＦＵと１−ＯＨＰの併用投与では、殆ど全ての投与条件で５−ＦＵの投与量や投与方法に関係なく相乗効果が得られ、シスプラチンの場合と同様に条件によっては期待値の２倍の腫瘍細胞の死滅効果が現れる。検索した併用剤のうち、最も優れた結果が得られ、１−ＯＨＰとの併用剤としては５−ＦＵが最も推奨される。
【００１６】
イリノテカンと１−ＯＨＰの併用では、１−ＯＨＰを追加投与する全ての組合せで相乗効果が現れる。
アドリアマイシンと１−ＯＨＰの併用では、１−ＯＨＰを追加投与する殆どの組合せで相乗効果が現れる。
エトポシドと１−ＯＨＰの併用では、１−ＯＨＰを追加投与する殆どの組合せで相乗効果が現れる。
マイトマイシンと１−ＯＨＰの併用では、１−ＯＨＰを先行投与する多くの組合せで相乗効果が現れる。
ミトキサントロンと１−ＯＨＰの併用では、少量のミトキサントロンを使用しかつ１−ＯＨＰを追加投与する全ての組合せで相乗効果が現れる。
ブレオマイシンと１−ＯＨＰの併用では、１−ＯＨＰを第３日又は第４日に追加投与する多くの組合せで相乗効果が現れる。
又カルボプラチン、テガフール、カルモフール、ドキシフルリジン及びウラシルについては、シスプラチンや５−ＦＵと同等又は若干劣る効果が得られると期待される。５−ＦＵ及びその誘導体から成る単剤又は配合剤（テガフールとウラシルの混合）と１−ＯＨＰとの併用効果も、５−ＦＵと１−ＯＨＰの併用効果と同様に期待できる。
【００１７】
これらの効果的な併用剤は単独で１−ＯＨＰと併用するだけでなく、２種類以上の併用剤、例えば５−ＦＵとシスプラチンを１−ＯＨＰと併用するとより一層の効果が期待できる。
ビンクリスチンと１−ＯＨＰの併用では、相乗効果を示す組合せは全くなく、多くの組合せが相加効果を示す。
ビンデシンと１−ＯＨＰの併用では、１４種類の組合せで相乗効果を示す組合せは全くなく、相加効果を示すものが僅か３例であり、検索した併用剤のうち、最も劣っている。
【００１８】
前述した通り併用剤の種類だけでなく、投与量の大小、及び投与時期も併用効果に影響を及ぼす。投与量については明確な基準はないが、投与時期は各併用剤によって効果に対して明確な影響を及ぼす。例えばシスプラチンでは、１−ＯＨＰを先行投与しシスプラチンを追加投与する殆どの場合に期待値の約２倍の効果が現れるのに対し、シスプラチンを先行投与する場合には６種類の組合せ中１種類のみに相乗効果を示し、３種類の組合せでは相乗効果以下であった。
【００１９】
患者に対して薬剤を投与する場合、時間差投与より同時投与の方が、患者及び医師の負担が軽くなるため好ましいが、５−ＦＵは同時投与で期待値の１８０％又は１５０％の効果が生じ、この点からも５−ＦＵと１−ＯＨＰとが好ましい組合せであることが分かる。なお本発明の同時投与とは、複数の抗癌剤を時間差零で同時に投与する場合と、複数の抗癌剤を別個の投与操作により連続的に投与する僅かな時間差がある場合の両者を含み、これら以外の投与は時間差投与である。
本発明の下記実施例では、白血病の腫瘍細胞に対してのみ、本発明の併用投与を行なったが、他の臓器癌に関しても本発明方法及び本発明の混合抗癌剤は相乗効果又は相加効果を発揮することが期待される。
又抗癌剤の投与経路は、従来の通り経口、注射、膣又は肛門からの投与、及び皮膚への塗布等がある。そして前述した１−ＯＨＰと併用剤を同時投与することにより効果が生ずる組合せの場合には、両薬剤を混合して錠剤化したり、注射用アンプル中に封入したりすることができる。又混合する以外に、化学結合により１−ＯＨＰを併用剤と一体化して錠剤中に添加したりアンプル中に封入したりすることができる。時間差投与が望ましい場合には、１−ＯＨＰと併用剤の両者を別個に準備し、いずれか一方を先行投与し、他方を追加投与する。
なお次に記載する実施例における抗癌剤併用の評価方法は、本発明以外の抗癌剤併用にも適用でき、今後の抗癌剤併用の評価方法の指標となりうる有用な方法である。
【００２０】
【実施例】
次に本発明に係わる抗癌剤の併用投与に関する実施例を説明するが、本実施例は本発明を限定するものではない。
対象とした腫瘍細胞はヒト白血球細胞株化細胞（ＲＰＭＩ−８４０２、ギブコ社製）であり、該細胞に１５％の牛胎児血清（ギブコ社製）を加えて細胞数を１０⁵ 個／mlとした細胞浮遊液を５mlの培養瓶（商品名スミロン、住友ベークライト株式会社製）中に調製した。この培養瓶中に調製した浮遊液は、同一のものを多数準備し、併用剤としてシスプラチン、５−フルオロウラシル、イリノテカン、アドリアマイシン、エトポシド、マイトマイシン、ミトキサントロン、ブレオマイシン、ビンクリスチン及びビンデシンの中の１種類を選定し、次の各実験を別個に行なった。なお対照〔薬剤添加なし、１−ＯＨＰのみの添加、併用剤のみの添加、下記の項目（１）〜（４）〕、及び併用剤と１−ＯＨＰの併用投与の時間的経過を示すと図１のようになる。
【００２１】
（１）対照であり、薬剤は添加しなかった。
（２）１−ＯＨＰのみを第１日（ｄａｙ１）に添加した。
（３）多量の併用剤を第１日に添加した（シスプラチンは１μｇ／ml、５−ＦＵは１０μｇ／ml、アドリアマイシンは０．０５μｇ／ml、エトポシドは１μｇ／ml、そして比較としてビンクリスチン０．０５μｇ／ml、及びビンデシン０．０５μｇ／ml）。なおイリノテカンについてはイリノテカンの添加量を１０μｇ／mlに固定し、１−ＯＨＰの添加量を５μｇ／mlとした。
（４）少量の併用剤を第１日に添加した（シスプラチンは０．５μｇ／ml、５−ＦＵは５μｇ／ml、アドリアマイシンは０．０１μｇ／ml、エトポシドは０．５μｇ／ml、そして比較としてビンクリスチン０．０１μｇ／ml、及びビンデシン０．０１μｇ／ml）。なおイリノテカンについてはイリノテカンの添加量を１０μｇ／mlに固定し、１−ＯＨＰの添加量を１μｇ／mlとした。
【００２２】
（５）１−ＯＨＰと多量の併用剤〔項目（３）と同一量、以下同じ〕を同時に第１日に添加した。
（６）１−ＯＨＰと少量の併用剤〔項目（４）と同一量、以下同じ〕を同時に第１日に添加した。
（７）１−ＯＨＰを第１日に添加し、多量の併用剤を第２日に添加した。
（８）１−ＯＨＰを第１日に添加し、少量の併用剤を第２日に添加した。
（９）１−ＯＨＰを第１日に添加し、多量の併用剤を第３日に添加した。
（１０）１−ＯＨＰを第１日に添加し、少量の併用剤を第３日に添加した。
【００２３】
（１１）１−ＯＨＰを第１日に添加し、多量の併用剤を第４日に添加した。
（１２）１−ＯＨＰを第１日に添加し、少量の併用剤を第４日に添加した。
（１３）多量の併用剤を第１日に添加し、１−ＯＨＰを第２日に添加した。
（１４）少量の併用剤を第１日に添加し、１−ＯＨＰを第２日に添加した。
（１５）多量の併用剤を第１日に添加し、１−ＯＨＰを第３日に添加した。
（１６）少量の併用剤を第１日に添加し、１−ＯＨＰを第３日に添加した。
（１７）多量の併用剤を第１日に添加し、１−ＯＨＰを第４日に添加した。
（１８）少量の併用剤を第１日に添加し、１−ＯＨＰを第４日に添加した。
【００２４】
これらの各実験において、各浮遊液に５％炭酸ガスを加え、３７℃で６日間培養し、最初の薬剤投与（第１日）から、２４時間後（第２日）、４８時間後（第３日）、７２時間後（第４日）、９６時間後（第５日）、１２０時間後（第６日）、１４４時間後（第７日）における細胞の一定個数（１０００個）当たりの死細胞個数を測定した。該測定は培養液から試料をサンプリングし、細胞観察板（エレコン化学株式会社製ＺＯＧ−１）に入れ、１５００倍の蛍光装置付き倒立顕微鏡（ニコン株式会社製）で細胞を観察し、細胞１０００個当たりの死細胞数をカウントして行なった。死細胞数のカウントは１回の実験で３回観察し、その百分比と標準偏差値を算出した。
【００２５】
これらの測定値から、併用効率ＯＥ（期待値と実測値の比）を次の式を使用して算出した。
ＯＥ（％）＝Ａ／〔Ｃ₀ ＋（Ｂ−Ｃ₁ ）＋（Ｄ−Ｃ₂ ）〕×１００ （１）
式中、Ａは併用によって得られた第７日の死細胞の実測値、Ｃ₀ は上記（１）の対照細胞での第７日の殺細胞値、Ｂは１−ＯＨＰに曝されていた日数間の殺細胞効果（％）、Ｃ₁ はＢと同日数での上記（２）の対照細胞の殺細胞効果（％）、Ｄは併用剤に曝されていた日数間の殺細胞効果（％）、Ｃ₂ はＤと同日数での上記（３）の対照細胞の殺細胞効果（％）である。
【００２６】
各併用剤ごとの算出値を纏めると図２の通りである。
図中、●、▲、および「黒正方形」（すなわち黒ぬり）は併用剤多量投与、○、△および□（すなわち白ぬき）は併用剤少量投与、●、および○（すなわち円印）は同時投与、▲、および△（すなわち三角）は１−ＯＨＰ先行投与、「黒正方形」および□（すなわち正方形）は併用剤先行投与、添数字２〜４は追加薬剤の投与日である。
図２から、各併用剤について前述した通りの効果が観察される。この効果を纏めると表１の通りになる。表中●は相乗効果を、○は相加効果を、△は相加効果未満であることをそれぞれ示している。
【００２７】
【表１】

【００２８】
【発明の効果】
本発明は、イリノテカン、及び所望によりさらにシスプラチン、カルボプラチン、５−フルオロウラシル、テガフール、カルモフール、ドキシフルリジン、ウラシル、アドリアマイシン、エトポシド、マイトマイシン、ミトキサントロン及びブレオマイシンから選択される１又は２以上の併用剤を、シス−オキザラート（１Ｒ，２Ｒ−ジアミノシクロヘキサン）白金（II）と併用投与することを特徴とする抗癌剤の併用投与方法である。
実施例から明らかなように、１−ＯＨＰとイリノテカン、そして所望によりさらに既存の抗癌剤を組合せて腫瘍細胞に投与すると、その殆ど全ての場合に該腫瘍細胞の死滅に対する効果が１−ＯＨＰ単独の場合に対して相加効果を有し、又多くの場合に相乗効果を生じさせる。
【００２９】
抗癌剤の併用投与による腫瘍細胞の死滅効果の増大は、特に腫瘍細胞の増殖速度と拮抗する近傍において重要な意味を持ち、僅かな死滅効果の増大でもその効果が重大な治療効果の促進をもたらすことになる。
本発明の抗癌剤の併用投与方法によると、特にその抗癌剤の組合せを適切に選択することによる死滅効果の増大は飛躍的であり、多くの癌治療に対して多大な貢献を行ない得る。
【００３０】
本発明方法の実施に使用する併用抗癌剤は、イリノテカン、さらには所望によりシスプラチン、カルボプラチン、５−ＦＵ、テガフール、カルモフール、ドキシフルリジン、ウラシル、アドリアマイシン、エトポシド、マイトマイシン、ミトキサントロン及びブレオマイシンから選択される１又は２以上の併用剤と１−ＯＨＰの組合せであり、該併用抗癌剤の使用により、前述した顕著な癌治療効果が生ずる。
【図面の簡単な説明】
【図１】対照、及び併用剤と１−ＯＨＰの併用投与の時間的経過を示すダイアグラム。
【図２】実施例における併用剤と１−ＯＨＰの併用投与の各条件における死滅効果の算出値を示すダイアグラム。[0001]
[Industrial application fields]
The present invention relates to a concomitant agent having a synergistic effect of an anticancer agent, and more specifically, a concomitant agent of cis-oxalate (1R, 2R-diaminocyclohexane) platinum (II) and irinotecan, which comprises cis-oxalate (1R , 2R-diaminocyclohexane) platinum (II) prior to administration of irinotecan, relates to a combined anticancer agent.
[0002]
[Prior art and problems]
Anticancer agents are generally used in combination with other anticancer agents in order to increase their effectiveness and prevent tumor cells from acquiring resistance to the drug. However, there is no special method for selecting a drug to be used in combination, which is a general condition as a drug to be used in combination, (1) that the drug used in combination is effective even when used alone, and (2) there are side effects. Under the general condition of not overlapping with concomitant drugs, concomitant drugs have been selected as a result of clinician experience.
[0003]
Tumor cells die when a reaction that constantly seeks to recover from damage caused by drugs (anticancer drugs) is induced in the cells, and the repair mechanism is not fully exerted. However, it takes time to die (necessity of time administration). When two or more types of anticancer agents are used in combination, cell biological reactions (synergistic effects) that cannot be observed by single administration may be exhibited. In this case, the specific mechanism of action possessed by each anticancer agent (all elucidated) May appear as competing or antagonistic reactions. In order to elucidate these, it is necessary to carry out various administrations combining the doses and administration times of the concomitant drugs.
[0004]
The present inventor has developed a rational technique for the combined use of anticancer agents (CANCER CHEMOTHERAPY: Challenges for the Future, vol.4, 1989: Molecular Bilology of DNA Topoisomerases, Proceedings of the International Symposium on DNA Topoisomerases in Chemotherapy, 1991) The effect of the combined use of various anticancer agents was demonstrated. However, there are few combinations of anticancer agents that show a synergistic effect, and a satisfactory combination effect of anticancer agents cannot be obtained easily.
[0005]
OBJECT OF THE INVENTION
The present invention has been made to solve these drawbacks. A novel anticancer agent, cis-oxalate (1R, 2R-diaminocyclohexane) platinum (II) (hereinafter referred to as 1-OHP), a known anticancer agent, Various studies have been conducted on the combination administration of these drugs, and an optimal combination administration method has been found and the present invention has been achieved.
[0006]
[Means for solving problems]
The present invention relates to irinotecan and optionally further selected from cisplatin, carboplatin, 5-fluorouracil (hereinafter also referred to as 5-FU), tegafur, carmofur, doxyfluridine, uracil, adriamycin, etoposide, mitomycin, mitoxantrone and bleomycin. Alternatively, two or more concomitant drugs can be combined with 1-OHP, and can be adjusted by mixing 1-OHP with a method of concomitant administration of an anticancer agent, a concomitant anticancer agent that can be used in the method, and 1-OHP. It is a mixed anticancer agent.
[0007]
Details of the present invention will be described below.
Tumor cells (cancer cells) have a higher growth rate than normal cells, and if the killing effect of the tumor cells is equal to or less than the growth rate, the progression of the cancer is suppressed and cannot be a fundamental treatment for cancer. . Each anticancer agent has an optimum dose, and even if a larger amount of anticancer agent is administered, the killing effect of tumor cells does not increase proportionally, but increases only slightly. Moreover, adverse effects such as damage of normal cells due to large doses often appear more strongly, and an increase in therapeutic effect due to large doses of a single anticancer drug can hardly be expected.
[0008]
Conventional administration of an anticancer agent in combination eliminates the limitations of using a single anticancer agent in a large amount and expands the possibility of cancer treatment. Although the mechanism of action has not been fully elucidated, it has been pointed out that the use of specific combinations may significantly increase the killing effect of tumor cells. In other words, by using anticancer agent A and anticancer agent B, an effect equal to or greater than the sum (additive effect) of the tumor cell killing rate when using anticancer agent A alone and the tumor cell killing rate when using anticancer agent B alone (additional effect) Synergy effect) may appear.
[0009]
Anticancer agents are not sufficient to suppress the progression of cancer, and it is desirable to completely kill tumor cells. For that purpose, it is necessary to kill the tumor cell more than the growth rate of the tumor cell. Theoretically, if the death rate of the tumor cell by the anticancer agent is slightly higher than the growth rate, the tumor cell will eventually be completely killed. To do. However, due to the risk of tumor cell metastasis and the risk of administering a long-term toxic anticancer agent to a patient, the emergence of anticancer agents capable of killing tumor cells in a short period of time is desired. For this purpose, a combination of anticancer agents that synergistically increase the killing effect of tumor cells by using two or more anticancer agents is essential, and the emergence of this combination of anticancer agents having a synergistic effect is decisive for current cancer chemotherapy. Contribute.
[0010]
In particular, the present invention proposes a combination of anticancer agents having at least an additive effect with this synergistic effect. 1-OHP represented by (Chemical Formula 1) is used as a target anticancer agent, and 1-OHP and 1 or 2 are combined. A combination of the above known anticancer agents is administered in combination. By this combined administration, the anticancer effect of 1-OHP is improved synergistically or additively, and a great contribution to cancer treatment can be expected by the combined administration.
[0011]
[Chemical 1]

[0012]
The present inventors have developed a total of 15 known anticancer agents (combination agents) including cisplatin, carboplatin, 5-FU, tegafur, carmofur, doxyfluridine, uracil, irinotecan, adriamycin, etoposide, mitomycin, mitoxantrone, bleomycin, vincristine and vindesine. ) And 1-OHP (targeting agent) in combination, the tumor cell killing effect is measured. In the case of the combined use, not only the combined use of anticancer agents but also the administration method (simultaneous administration, additional administration) and the dosage (large dose, small dose) are examined.
[0013]
In evaluating the killing effect of the tumor cells in each combination, the killing effect of the control (when no anticancer agent is used) is subtracted, and the killing effect by the combined use of the pure combination agent and 1-OHP is calculated.
The combined effect is calculated by using the following formula (1), and when the value (%) calculated by this formula is 85 to 115%, there is an additive effect, and when the value is less than 85% Is considered to have a synergistic effect if it exceeds 115% below the additive effect. ± 15% is a confidence limit or error.
[0014]
As a result, as described in detail below, most of the eight types of cisplatin, 5-FU, irinotecan, adriamycin, etoposide, mitomycin, mitoxantrone, and bleomycin are additive regardless of the administration method and dose. An effect or a synergistic effect can be obtained, particularly in the combination of cisplatin and 5-FU, and the combination of 5-FU and 1-OHP is the best. On the other hand, there are many combinations of vincristine and vindesine that only have an additive effect at best.
[0015]
That is, in the combined administration of cisplatin and 1-OHP, a synergistic effect is obtained when cisplatin is coadministered and all combinations where 1-OHP is administered prior to cisplatin, and the expected value (additive effect) may be obtained depending on conditions. Twice the tumor cell killing effect appears.
In the combined administration of 5-FU and 1-OHP, a synergistic effect is obtained regardless of the dose and administration method of 5-FU under almost all administration conditions, and double the expected value depending on the conditions as in the case of cisplatin. The tumor cell killing effect appears. Among the searched concomitant drugs, the best results are obtained, and 5-FU is most recommended as a concomitant drug with 1-OHP.
[0016]
In the combined use of irinotecan and 1-OHP, a synergistic effect appears in all combinations where 1-OHP is additionally administered.
In the combination of adriamycin and 1-OHP, a synergistic effect appears in most combinations where 1-OHP is additionally administered.
With the combination of etoposide and 1-OHP, a synergistic effect appears in most combinations where 1-OHP is additionally administered.
In the combination of mitomycin and 1-OHP, a synergistic effect appears in many combinations where 1-OHP is administered in advance.
In the combination of mitoxantrone and 1-OHP, a synergistic effect appears in all combinations where a small amount of mitoxantrone is used and 1-OHP is additionally administered.
In the combination of bleomycin and 1-OHP, a synergistic effect appears in many combinations where 1-OHP is additionally administered on the third or fourth day.
Carboplatin, tegafur, carmofur, doxyfluridine and uracil are expected to have the same or slightly inferior effects as cisplatin and 5-FU. The combined effect of 1-OHP with a single agent or compounding agent (mixture of tegafur and uracil) composed of 5-FU and its derivatives can be expected similarly to the combined effect of 5-FU and 1-OHP.
[0017]
These effective concomitant drugs are not only used in combination with 1-OHP alone, but also more effects can be expected when two or more kinds of concomitant drugs, for example, 5-FU and cisplatin are used in combination with 1-OHP.
In the combination of vincristine and 1-OHP, there is no combination showing a synergistic effect, and many combinations show an additive effect.
In the combined use of vindesine and 1-OHP, there is no combination that shows a synergistic effect among the 14 types of combinations, and only 3 cases show an additive effect, and the worst combination among the searched concomitant drugs.
[0018]
As described above, not only the type of concomitant drug but also the size and timing of administration affect the concomitant effect. There is no clear standard for the dose, but the timing of administration has a clear effect on the effect of each concomitant drug. For example, in the case of cisplatin, the effect of about twice the expected value appears in most cases where 1-OHP is administered in advance and cisplatin is additionally administered, whereas when cisplatin is administered in advance, only one of the 6 combinations is available. A synergistic effect was exhibited in the three combinations, which were less than the synergistic effect.
[0019]
When administering drugs to patients, simultaneous administration is preferable to time difference administration because it reduces the burden on the patient and doctor, but 5-FU produces an effect of 180% or 150% of the expected value by simultaneous administration. From this point, it can be seen that 5-FU and 1-OHP are a preferred combination. The simultaneous administration of the present invention includes both a case where a plurality of anticancer agents are administered simultaneously with zero time difference and a case where there is a slight time difference in which a plurality of anticancer agents are continuously administered by separate administration operations. Administration is time lag.
In the following examples of the present invention, the combined administration of the present invention was performed only on leukemia tumor cells, but the method of the present invention and the mixed anticancer agent of the present invention also exhibited synergistic effects or additive effects for other organ cancers. Expected to demonstrate.
As for the administration route of the anticancer agent, there are conventional methods such as oral administration, injection, administration from the vagina or anus, and application to the skin. In the case of a combination that produces an effect by simultaneously administering 1-OHP and a concomitant drug, the two drugs can be mixed and tableted, or enclosed in an ampule for injection. In addition to mixing, 1-OHP can be integrated with a concomitant agent by chemical bonding and added to a tablet or enclosed in an ampoule. When time-lag administration is desired, both 1-OHP and the concomitant drug are prepared separately, one of which is administered in advance and the other is administered additionally.
In addition, the evaluation method of the anticancer agent combination in the Example described below can be applied to the anticancer agent combination other than the present invention, and is a useful method that can serve as an index of a future evaluation method of the anticancer agent combination.
[0020]
【Example】
Next, examples relating to the combined administration of anticancer agents according to the present invention will be described, but the present examples are not intended to limit the present invention.
The target tumor cells were human leukocyte cell lines (RPMI-8402, manufactured by Gibco), and 15% fetal bovine serum (Gibco) was added to the cells to make the number of cells 10 ⁵ / ml. The cell suspension was prepared in a 5 ml culture bottle (trade name Sumilon, manufactured by Sumitomo Bakelite Co., Ltd.). A large number of the same suspensions prepared in this culture bottle are prepared, and one kind of cisplatin, 5-fluorouracil, irinotecan, adriamycin, etoposide, mitomycin, mitoxantrone, bleomycin, vincristine and vindesine is used as a concomitant agent. And the following experiments were performed separately. The time course of the control [no addition of drug, addition of 1-OHP only, addition of concomitant only, items (1) to (4) below] and concomitant administration of 1-OHP with concomitant is shown. It becomes like 1.
[0021]
(1) Control, no drug added.
(2) Only 1-OHP was added on the first day (day 1).
(3) A large amount of concomitant drug was added on the first day (cisplatin 1 μg / ml, 5-FU 10 μg / ml, adriamycin 0.05 μg / ml, etoposide 1 μg / ml, and vincristine 0.05 μg as a comparison. / Ml, and vindesine 0.05 μg / ml). For irinotecan, the amount of irinotecan added was fixed at 10 μg / ml, and the amount of 1-OHP added was 5 μg / ml.
(4) A small amount of concomitant drug was added on day 1 (cisplatin 0.5 μg / ml, 5-FU 5 μg / ml, adriamycin 0.01 μg / ml, etoposide 0.5 μg / ml, and for comparison Vincristine 0.01 μg / ml and vindesine 0.01 μg / ml). For irinotecan, the amount of irinotecan added was fixed at 10 μg / ml, and the amount of 1-OHP added was 1 μg / ml.
[0022]
(5) 1-OHP and a large amount of a concomitant agent [the same amount as item (3), the same applies hereinafter) were simultaneously added on the first day.
(6) 1-OHP and a small amount of a concomitant agent [the same amount as item (4), the same applies hereinafter) were simultaneously added on the first day.
(7) 1-OHP was added on the first day, and a large amount of the concomitant agent was added on the second day.
(8) 1-OHP was added on the first day, and a small amount of the concomitant agent was added on the second day.
(9) 1-OHP was added on the first day, and a large amount of the concomitant drug was added on the third day.
(10) 1-OHP was added on day 1 and a small amount of concomitant was added on day 3.
[0023]
(11) 1-OHP was added on the first day, and a large amount of the concomitant agent was added on the fourth day.
(12) 1-OHP was added on day 1 and a small amount of concomitant was added on day 4.
(13) A large amount of the concomitant agent was added on the first day, and 1-OHP was added on the second day.
(14) A small amount of the concomitant agent was added on the first day, and 1-OHP was added on the second day.
(15) A large amount of the concomitant drug was added on the first day, and 1-OHP was added on the third day.
(16) A small amount of the concomitant agent was added on the first day, and 1-OHP was added on the third day.
(17) A large amount of the concomitant drug was added on the first day, and 1-OHP was added on the fourth day.
(18) A small amount of the concomitant agent was added on the first day, and 1-OHP was added on the fourth day.
[0024]
In each of these experiments, 5% carbon dioxide gas was added to each suspension and cultured at 37 ° C. for 6 days. After the first drug administration (Day 1), 24 hours (Day 2), 48 hours (Day 2) 3 days), 72 hours (4th day), 96 hours (5th day), 120 hours (6th day), 144 hours (7th day) per certain number of cells (1000 cells) The number of dead cells was measured. In this measurement, a sample is sampled from the culture solution, placed in a cell observation plate (ZOG-1 manufactured by Elecon Chemical Co., Ltd.), and observed with an inverted microscope with a 1500 × fluorescent device (manufactured by Nikon Co., Ltd.). The number of dead cells per count was counted. Counting the number of dead cells was observed three times in one experiment, and the percentage and standard deviation value were calculated.
[0025]
From these measured values, combined use efficiency OE (ratio of expected value and actual value) was calculated using the following equation.
OE (%) = A / [C ₀ + (B−C ₁ ) + (D−C ₂ )] × 100 (1)
In the formula, A is the actual value of the dead cell on the 7th day obtained by the combination, C ₀ is the cell killing value on the 7th day in the control cell of (1) above, and B is exposed to 1-OHP. cytotoxic effect between days (%), C ₁ is the cellular effects (%) killing of control cells (2) in the B and the number of days, D is between days that were exposed to the combination agent cytotoxic effect ( %), C ₂ is the cell killing effect (%) of the control cells of (3) above in the same number of days as D.
[0026]
The calculated values for each concomitant drug are summarized as shown in FIG.
In the figure, ●, ▲, and “black square” (that is, blackening) are a large amount of concomitant drug administration, ○, △, and □ (that is, whitening) are a small amount of concomitant drug administration, and ● and ○ (that is, a circle) are simultaneous Administration, ▲, and △ (ie, triangles) are 1-OHP prior administration, “black squares” and □ (ie, squares) are concomitant medication prior administrations, and additional numbers 2 to 4 are the dates of administration of additional agents.
From FIG. 2, the effects as described above are observed for each combination drug. Table 1 summarizes this effect. In the table, ● indicates a synergistic effect, ○ indicates an additive effect, and △ indicates that the additive effect is less than the additive effect.
[0027]
[Table 1]

[0028]
【The invention's effect】
The present invention relates to irinotecan and optionally one or more combinations selected from cisplatin, carboplatin, 5-fluorouracil, tegafur, carmofur, doxyfluridine, uracil, adriamycin, etoposide, mitomycin, mitoxantrone and bleomycin, It is a combined administration method of an anticancer agent, characterized by being administered in combination with cis-oxalate (1R, 2R-diaminocyclohexane) platinum (II).
As is apparent from the Examples, when 1-OHP and irinotecan, and optionally an existing anticancer agent, are administered to tumor cells in combination, the effect on the killing of the tumor cells is almost always 1-OHP alone. Have an additive effect and often a synergistic effect.
[0029]
Increasing the killing effect of tumor cells by the combined administration of anticancer drugs is important especially in the vicinity of antagonizing the growth rate of tumor cells, and even if a slight increase in killing effect, the effect leads to the promotion of a significant therapeutic effect become.
According to the combined administration method of the anticancer agent of the present invention, the increase in the killing effect by selecting an appropriate combination of the anticancer agents is dramatic, and can make a great contribution to many cancer treatments.
[0030]
The combined anticancer agent used in the practice of the method of the present invention is irinotecan, and optionally 1 selected from cisplatin, carboplatin, 5-FU, tegafur, carmofur, doxyfluridine, uracil, adriamycin, etoposide, mitomycin, mitoxantrone and bleomycin Or it is a combination of two or more concomitant drugs and 1-OHP, and the use of the concomitant anticancer drug produces the remarkable cancer therapeutic effect described above.
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a diagram showing the time course of control and the combined administration of 1-OHP with a combination drug.
FIG. 2 is a diagram showing a calculated value of a killing effect under each condition of combined administration of a concomitant drug and 1-OHP in Examples.

Claims (3)

Cis - oxalate (1R, 2R- diaminocyclohexane) a combination drug of the platinum (II) and irinotecan, cis - oxalate (1R, 2R- diaminocyclohexane) irinotecan prior to administration of platinum (II) is administered , Combination anticancer drugs. The combined anticancer agent according to claim 1, wherein at least one of cisplatin, carboplatin, 5-fluorouracil, degafur, carmofur, doxyfluoridine, uracil, adriamycin, etoposide, mitomycin, mitoxantrone or bleomycin is used as a further component. .   The combination anticancer agent according to claim 2, wherein the further component is administered after administration of cis-oxalate (1R, 2R-diaminocyclohexane) platinum (II).

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